1. Field of the Invention
The present invention relates to a method for immobilizing glycoprotein, and more particularly to a method for immobilizing glycoprotein by forming covalent bonding with two spatially adjacent hydroxyl groups of a carbohydrate chain.
2. Description of the Prior Art
In the development of protein microarray, the accessibility of surface protein active site and stability may be significantly influenced by its orientation on the solid surface. Therefore, various site-specific immobilization strategies have been developed.
Due to tight and specific epitope affinities, antibodies have been widely used in immunoaffinity extraction, targeted therapeutic delivery, enzyme-linked immunosorbent assays, and diagnostic arrays.
Traditionally, antibodies have been modified at lysine, arginine, aspartate, and glutamate residues by random amide bond conjugation or Schiff base formation. However, such random conjugation often reduces antibody function to a point that is inadequate for the desired application.
In addition, antibody binding proteins, e.g. protein G and protein A, are commonly used to anchor/immobilize antibody via the Fc (Fragment, crystallizable) region in a site-specific manner for exposing the Fab (Fragment, antigen-binding) region in an efficient way. However, the aforementioned non-covalent coupling between the antibody binding protein and antibody is likely subjected to dissociation caused by environmental factors such as pH value, salinity, and temperature.
Therefore, it is a now current goal to develop a method for immobilizing glycoprotein which is site-specific and covalent protein immobilization without interfering the binding affinity of an antibody to antigen.